Human INA (alpha Internexin) knockout HeLa cell lysate (ab257482)
Overview
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Product name
Human INA (alpha Internexin) knockout HeLa cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 179 bp deletion in exon1 and 1 bp insertion in exon1. -
Passage number
Knockout validation
Sanger Sequencing, Western Blot (WB)Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262102 - Human INA knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Target
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Function
Class-IV neuronal intermediate filament that is able to self-assemble. It is involved in the morphogenesis of neurons. It may form an independent structural network without the involvement of other neurofilaments or it may cooperate with NF-L to form the filamentous backbone to which NF-M and NF-H attach to form the cross-bridges. -
Tissue specificity
Found predominantly in adult CNS. -
Sequence similarities
Belongs to the intermediate filament family. -
Developmental stage
Expressed in brain as early as the 16th week of gestation, and increased rapidly and reached a steady state level by the 18th week of gestation. -
Post-translational
modificationsO-glycosylated.
Phosphorylated upon DNA damage, probably by ATM or ATR. - Information by UniProt
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Alternative names
- 66 kDa neurofilament protein
- AINX_HUMAN
- Alpha Inx
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262102 - Human INA knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Images
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Western blot - Human INA knockout HeLa cell lysate (ab257482)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: INA knockout HeLa cell lysate (20 µg)
Lane 3: SH-SY5Y cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab40758 observed at 66 kDa. Red - loading control ab8245 observed at 36 kDa.
ab40758 Anti-alpha Internexin antibody [EP676Y] was shown to specifically react with alpha Internexin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265264 (knockout cell lysate ab257482) was used. Wild-type and alpha Internexin knockout samples were subjected to SDS-PAGE. ab40758 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human INA knockout HeLa cell lysate (ab257482)Allele-1: 1 bp insertion in exon1
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Sanger Sequencing - Human INA knockout HeLa cell lysate (ab257482)Allele-2: 179 bp deletion in exon1
